Abstract To evaluate the therapeutic applications of stem cells, it is essential to develop translational research using several animal species. Rabbit is a useful model for biomedical research. It has been used to study a number of human diseases. Compared to mice, rabbits have a more similar embryology and stem cell biology to those of humans, an adult body size comparable to human infants that allows for practical surgical procedures, and a longer lifespan to permit a clinically relevant research time. It will be beneficial to establish rabbit pluripotent stem cell (PSC) lines as a non-murine model system for better understanding human stem cells, and for testing the safety and efficacy of new stem cell based technologies. The availability of ground state rabbit ESCs in the age of gene editing is expected to create many novel research tools for translational research community. Pluripotent rabbit embryonic stem cells (rbESCs) and induced pluripotent stem cells (iPSCs) have been reported. However, these cells share typical properties of primed stem cells, and all failed to transmit to the germ cells. The ground state or germline transmitting ESCs and iPSCs are only reported in mice and rats. It remains to be tested whether ground state ESCs exist in non-rodent species. The PI of this project made the breakthrough in establishing the first non-murine ground state ESCs in rats. Here we propose to derive ground state rbESCs. In Aim 1, we will establish an Oct4-?PE-GFP reporter cell line, which would express green florescent in ground state but not in prime state. In Aim 2, we will screen small molecule library to identify compounds that are beneficial for the ground state in rabbit ESCs. In Aim 3, we will derive new ground state rabbit ESCs.